All the fabrication processes including the final cleaning can be finished within 4 h. All nanostructured quartz surfaces exhibited contact angles higher than 155° with minimal water droplet adhesiveness and enhanced transparency (due to antireflectivity) over a broad spectral range from 350 to 900 nm. Furthermore, A2 pattern showed an enhanced antireflective effect that extends to the deep-UV range near 190 nm, which is a drawback region in conventional thin-film-coating approaches as a result of thermal damage. Because, by changing the conditions of successive RIE, the geometrical configurations of nanostructure arrays can be easily modified to meet specific needs, the newly developed fabrication method is expected to be applied in various optic and opto-electrical areas.Both self-cleanability and antireflectivity were achieved on quartz surfaces by forming heptadecafluoro-1,1,2,2-tetrahydrodecyltrichlorosilane self-assembled monolayer after fabrication of nanostructures with a mask-free method. By exposing polymethylmethacrylate spin-coated quartz plates to OPACS codes: 06.60.Ei; 81.65.Cf; 81.40.Vw. Numerous studies of surface nanostructures have been conducted to investigate enhancement of the properties of bulk materials to improve their selectivity, applicability, and effectiveness. During the past decades, the technological basis for nanofabrication has been developed by vigorous efforts to develop next-generation lithography for highly resolved patterns up to the industrial level of semiconductor production. Nanofabrication techniques using transparent materials such as quartz comprise one of the most attractive approaches to optical and opto-electrical studies as well as to highly sensitive biosensor fields since quartz is commonly employed in these fields -4.Various methods are used to achieve antireflective property in quartz or glass to increase light transmission. Single- or multilayered thin film coatings, porous coatings, and fabrication of sub-wavelength nanostructures on surfaces using conventional lithography have been the focus of many studies -7. Howev2 and CF4 RIE) of polymer-coated quartz in our previous studies [In comparison with thin-film-coated surfaces, directly patterned surfaces usually guarantee good mechanical stability because they are free from adhesion problems and tensile stress. Recently, a few techniques for the direct fabrication of sub-wavelength nanostructures on quartz or glass surfaces have been attempted by several groups. Lohmüller et al. and Christopher et al. used block copolymer micelle lithography with reactive ion etching (RIE) and reported that array pattern of a quartz nanostructure showed excellent antireflectivity and anti-fogging in UV and deep-UV region ,9. Li et studies ,4.Superhydrophobic behavior of a surface can also be achieved by introducing micro- and/or nanostructures at the surface. A superhydrophobic surface is usually defined as having a contact angle greater than 150°. The self-cleaning effect refers to cases in which contaminant particles adhered to a surface are easily washed off with rolling of water droplets. To have a self-cleaning effect, the surface should possess minimized adhesion properties as well as superhydrophobicity -14.The mimetic fabrication of a superhydrophobic surface was primarily inspired by the self-cleanable leaves of the lotus, which have an array of protrusions on the surface ,16. The We previously reported a systematic approach to obtain a superhydrophobic surface with tunable adhesiveness and suggested a useful nanofabrication strategy for achieving self-cleanability that involved fabrication of pillar arrays without dead-end nanopores covered with low-surface-energy materials ,14. In tThe nanopillar arrays with various pillar diameters and inter-pillar distances were fabricated on quartz plates by a mask-free approach. A quartz wafer plate  was cleaned with piranha solution and then rinsed thoroughly with deionized (DI) water. After heating at 100°C for about 5 min, 950 PMMA A2, 495 PMMA A8, and 950 PMMA A11  were each spin-coated onto the surface at 4,000 rpm for 25 s. The expected thicknesses of the PMMA layers reported in the technical support information of the MicroChem products are 50, 500, and 800 nm for A2, A8, and A11, respectively.2 plasma for 1 min and CF4 plasma for 10 min at 250 W of RF power, 40 mTorr, and 40 sccm using a custom-made RIE system. To remove the organic remnants from the quartz pattern, each pattern was sintered at 1,000°C in a furnace for 1 h, cleaned with piranha solution, rinsed with DI water, and dried. The surface of each pattern was covered with a self-assembled heptadecafluoro-1,1,2,2-tetrahydrodecyltrichlorosilane  monolayer using a 3-mM solution of HDFS in n-hexane to reduce the surface energy. All the fabrication processes including the final cleaning and self-assembled monolayer formation could be finished within 4 h.Post-baking was conducted for 30 min at 170°C. The PMMA-coated quartz plates were then subjected to reactive ion etching in OThe morphological images of the nanopillars were obtained by field emission scanning electron microscopy . The surface-wetting properties were evaluated by an Easydrop goniometer , and the dynamic angle was evaluated using a DSA 100 goniometer (KRÜSS). The advancing and receding contact angles were obtained from more than three points in each specimen by the sessile drop technique. To investigate optical performance, we measured the transmission and reflection properties of the nanostructured quartz plates using a UV-visible spectrometer .2) in each sample. These results demonstrate that it is feasible to systematically control the dimensional features of the pillar pattern. Specifically, larger and higher pillars can be formed by controlling the thickness of the PMMA resist. The mechanism of the nanostructure formation was previously reported [2 and CF4 two-step RIE process because, by controlling the RIE conditions appropriately, CF4-resistant CxFy polymeric mask is automatically and selectively deposited during the CF4 RIE process on the top of the dot-like nanostructures of the PMMA resist, which are formed during the preceding O2 RIE process.Figure reported . ExplainTable d and the wavelength λ. For a graded-index transition, substantial antireflection can be obtained when the ratio d/λ is about 0.4 or higher [In nature, some insects have sub-wavelength scale structure patterns with nipple-like or tapered profiles on the cornea that exhibit a gradient in the refractive index between the air and tissue interface. These characteristics play an important role in increasing light transmission. Theoretically, for a thin-film coating, overall reflectance can be a function of antireflection (AR) layer thickness r higher ,10. To er higher .Figure 2. Third, our technology can be employed for industrial optical devices, optical components, and interior and exterior materials requiring both self-cleanable and antireflective properties. In addition, the manufacturing cost is minimal since the fabrication process is simple and fast, and requires no mask.The mask-free method presented here may have several advantages. First, long-term stability is expected because of the superhydrophobic self-cleaning effect. Second, although the area can be restricted by the stage size of a RIE device, a large area can be achieved because our method requires no masks. Figure In the near future, after proper optimization, we plan to apply our technique to other optical components, such as lenses and optical filters, to demonstrate the wide applicability of our technology. Furthermore, we will determine whether the antireflective spectral regions can be controlled using different PMMA resists by changing the parameters of the surface structures such as height, width, and pitch.The authors declare that they have no competing interests.JSK prepared the samples, evaluated surface characteristics, and drafted the manuscript. HWJ evaluated the optical properties, analyzed the data, and drafted the manuscript. WL participated in the design of the study and drafted the manuscript. BGP evaluated the surface characteristics of the samples. BMK and KBL conceived of the study together and participated in its design and coordination. All authors read and approved the final manuscript.Video S1. Video shows the rolling of water droplets on the self-cleanable nanostructured quartz surface prepared using PMMA A2, PMMA A8, and PMMA A11.Click here for file


In addition, the GTV was delineated using the signal-to-background (SBR) ratio-based adaptive thresholding technique (GTVSBR). Overlap analysis were conducted to assess geographic mismatches between the GTVs delineated using the different techniques. Inter- and intra-observer variability were also assessed.F-FDG PET/CT based RT treatment planning was performed in 10 patients with locally recurrent (n = 5) or post-surgical residual gynaecological cancer (n = 5). The gross tumor volume (GTV) was defined by 4 experienced radiation oncologists first using contrast enhanced CT (GTVCT (43.65 cm3) was larger than the mean GTVPET/CT (33.06 cm3), p = 0.02. In 6 patients, GTVPET/CT added substantial tumor extension outside the GTVCT even though 90.4% of the GTVPET/CT was included in the GTVCT and 30.2% of the GTVCT was found outside the GTVPET/CT. The inter- and intra-observer variability was not significantly reduced with the inclusion of 18F-FDG PET imaging . The GTVSBR was smaller than GTVCT p ≤ 0.005 and GTVPET/CT p ≤ 0.005.The mean GTV18F-FDG PET/CT images for target volume delineation of recurrent or post-surgical residual gynaecological cancer alters the GTV in the majority of patients compared to standard CT-definition. The use of SBR-based auto-delineation showed significantly smaller GTVs. The use of PET/CT based target volume delineation may improve the accuracy of RT treatment planning in gynaecologic cancer.The use of  In addition, the overlap volume of GTVPET/CT1 and GTVSBR relative to GTVSBR-OF was also calculated [OFSBR] (C).The delineated contours for both delineation phases were analyzed separately. Firstly, the volumes contoured by every observer for GTVInter- and intra-observer variability was calculated using a two-way ANOVA model. Regression analysis was used to evaluate the difference between calculated volumes and overlap between GTVs when using the different segmentation tools. Statistical analysis and curve fitting was performed using PASW Statistics package, version 18.0 . The level of statistical significance adopted was 0.05.18F-FDG PET/CT were able to pinpoint the local recurrent or residual cancer in the pelvis. The median SUVmax of GTVs was 11.74 . We did not observe any difference in PET signal between residual tumor and recurrent tumors. Figure CT and GTVPET/CT was observed. The mean GTVCT1  was significantly larger than the mean GTVPET/CT1 , p = 0.02. The smallest GTVCT1 and GTVPET/CT1 was found in patient #6 with 1.89 cm3 and 0.85 cm3 respectively, and the largest GTVCT in patient #4 with 120.39cm3, while the largest GTVPET/CT was observed in patient #10 (101.93 cm3). Figure The contrast enhanced CT scan as well as the 3 (SD 5.42) and the mean common volume was 31.48 cm3 (SD 4.21). The mean OFCT1 was 0.63 (SD 0.04). The mean OFPET/CT1 was 0.90 (SD 0.03). In 2 patients, the GTVPET/CT of all observers was included entirely in the GTVCT and in 6 patients, GTVPET/CT added substantial tumor extension outside the GTVCT.Table 18F-FDG-PET was included, this ratio was reduced to median 1.38 . The ratio of largest to smallest GTV was decreased in 9 of 10 patients using PET/CT for GTV delineation.We found that among four experienced radiation oncologists, the ratio of largest to smallest GTVs outlined on 10 patients using the planning CT had a median of 1.87 . When the CT was 0.37  and for the GTVPET/CT was 0.48 ; p = 0.23. All physicians contoured each patient twice and the median intra-observer percentage of concordance for the GTVCT was 0.49  and for the GTVPET/CT was 0.65  (p = 0.18).The median inter-observer reliability index for the GTVSBR are shown in Table SBR was 21.33 cm3 (SD 23.87), which is significantly smaller than the manually contoured GTVCT (p ≤ 0.005) and GTVPET/CT (p ≤ 0.005). In 6 patients the GTVSBR was included completely in all GTVCTs and the mean OF between GTVSBR and GTVPET/CT was 0.97 (SD 0.02). Comparing the GTVSBR with the GTVPET/CTs, we observe that in 4 patients the GTVSBR were larger than the GTVPET/CT.The GTVs were delineated both manually and by editing the SBR-based auto-contour. The results concerning GTVCT[18F-FDG PET/CT plays an increasingly important role in the staging and management of gynaecologic cancer including RT treatment planning [18F-FDG PET/CT has demonstrated a high sensitivity and accuracy of more than 90% with average specificity in locally advanced or recurrent gynaecologic pelvic carcinoma. Furthermore 18F-FDG PET/CT can help to distinguish between tumor recurrence and post-therapy changes [18F-FDG PET/CT-guided IMRT had improved survival and decreased treatment related toxicity compared with patients treated with non-IMRT radiotherapy [CT and MRI have reasonable sensitivity but low specificity in identifying recurrent gynaecologic disease ,2. ConseCT. 18F-FDGplanning ,4. 18F-F18F-FDG PET/CT-based target volume delineation in local recurrent or postsurgical residual gynaecological cancer. The results were compared with an automated PET segmented technique using adaptive thresholding technique. In other cancer sites such as head and neck and lung, 18F-FDG PET/CT was reported to decrease inter- and intra-observer variability in tumor contouring [18F-FDG PET/CT could be superior to CT alone in this group of patients. GTVPET/CT was significantly smaller than the GTVCT with a trend for reduced inter- and intra-observer variability using PET/CT. The inter-observer agreement was moderate for the GTVCT and substantial for the GTVPET/CT[18F-FDG PET/CT in our study is in line with these findings [18F-choline PET/CT. In gynaecologic cancer no inter- or intra-observer variability in PET-based GTV-delineation has been evaluated until now.This delineation study evaluated inter- and intra-observer variability of CT-based and ntouring . Our resGTVPET/CT. The intGTVPET/CT. It has findings . Only onfindings . A trendPET/CT was significantly smaller than the GTVCT with the implementation of a coregistered 18F-FDG PET/CT. When the GTVSBR volumes were analyzed and compared with manual delineated target volume, it was observed that the GTVSBR was significantly smaller than the median GTVCT and GTVPET/CT. This was also manifested in the overlap analysis, where the overlap fraction increased from OFCT1 to OFPET/CT1 and OFSBR. Overall, the comparison of GTVs delineated in primary and recurrent cancer did not result in any significant differences.Our study demonstrate that the size of GTVCT and GTVPET/CT determination and that the exams were performed on a dedicated PET/CT scanner for virtual simulation and fused with a recent MRI. Nevertheless the inter- and intra-observer variability was relatively high with both imaging modalities, highlighting the difficulty to determine the target volumes in this group of patients. An automated segmentation of the target volume using the adaptive thresholding technique could eventually help to reduce inter- and intra-observer variability. One potential limitation of our study is that the observers were at liberty to adjust the window, level and contrast setting of the images. This could have increased the inter- and intra-observer variability. However, all observers were experienced in PET/CT-based target volume delineation and were helped by a nuclear medicine physician. Another drawback of this study is the lack of comparison of the PET/CT results with pathologic findings after surgery.The strength of our study includes the use of contrast enhanced CT scans for GTThe delineation of target volumes and organ at risk is a very critical step in high-precision RT treatment planning. Good image quality and reliable delineation protocol are important for accurate target volume delineation. One of the challenges of PET/CT-guided target volume delineation is the accurate segmentation of noisy and low resolution functional PET images. This is in particular true in recurrent or residual gynaecological cancer where vascular and urinary activity hampers target volume delineation. The result is a relatively high inter- and intra-observer variability. Various PET image segmentation techniques for target volume delineation were developed and evaluated to overcome this drawback . Among tSBR in gynaecologic tumors is needed for validation purposes. PET-based target volume delineation in gynaecologic tumors is actually not recommended outside clinical studies. It has to be emphasized that both patients with recurrent or postsurgical residual gynaecologic cancer are challenging cohorts for reliable target volume delineation and thus it is more likely that high inter- and intra-observer variability will result. In the absence of a more accurate information on the target volume position in gynaecologic cancer, a composite of GTVCT and GTVPET/CT can be recommended to optimize the GTV definition.An improved concordance in target volume delineation using PET/CT implies a greater accuracy and can help to determine a more appropriate treatment plan. In our study, the inter-observer variability coefficient prevailed is still relatively high. Variability negatively impacts the quality of treatments delivered to cancer patients. Alternatively an automated segmented target volume could be considered. There is consensus in the need for highly objective and automatic segmentation methods, and various groups have observed that semi- or fully-automated delineation techniques reduce inter-observer variability and improve reproducibility . The adaPET/CT was significantly smaller than GTVCT. The reduction was larger when the adaptive thresholding-based semi-automated contouring algorithm was used. GTVPET/CT added substantial tumor extension outside the GTVCT in 60% of the patients. The combination of a matched 18F-FDG PET/CT reduced the inter- and intra-observer variation in the delineation of gynaecological cancer however the difference was not significant. Target volume delineation may be improved with the inclusion of 18F-FDG PET/CT.This delineation study showed that GTVThe authors declare that they have no competing interests.HV carried out the drafting of the manuscript and partially the analysis of the data. RM carried out the design of the study. HW and NC defined the target volumes and participated in the analysis of the data. TZ defined the target volumes and participated in the analysis of the data. YP participated in the target volume definition. HI performed the imaging analysis. HZ participated in the analysis of the data. OR participated in the methodological design. All authors read and approved the final manuscript.


Subsequently, life tables were constructed for assessment of radiation risks, also taking into account the disease-related mortality.in brackets] were 0.4% [0.6%] for boys and 0.7% [1.1%] for girls, and for adults with NHL 0.07% [0.28%] for men and 0.09% [0.37%] for women.In children with HD, cumulative effective dose from medical imaging ranged from 66 mSv (newborn) to 113 mSv (15 years old). In adults with NHL the cumulative effective dose from medical imaging was 97 mSv. Average fractions of radiation-induced deaths for children with HD [without correction for disease-related mortality 18F-FDG PET can be considered as justified, but imaging should be performed with care, especially in children.Taking into account the disease-related reduction in life expectancy of patients with malignant lymphoma results in a higher overall mortality but substantial lower incidence of radiation induced deaths. The modest radiation risk that results from imaging with CT and Survival of malignant lymphoma has improved dramatically over the past decades.PET and CT currently play important roles for malignant lymphoma patients.The potential hazard of ionising radiation has become an increasingly important issue.When assessing radiation risks, disease-related reduction in life expectancy should be considered.CT and18F-FDG PET create a modest radiation-induced mortality risk. Accurate imaging is important for determining the stage of disease, which guides the treatment strategy and influences the prognosis . Imaging imaging , 5–9. Honic risk –17.There is a worldwide growing concern about radiation exposure in medical imaging , 18–21. Therapeutic advances have dramatically improved the survival of patients with malignant lymphoma over the past decades. HD can nowadays be cured in at least 80% of patients –25. Ther18F-FDG PET was calculated for a typical work-up for children with HD in different age categories, and for adults with NHL. We expected that disease-related mortality would have a significant effect on the assessment of radiation risk in patients with malignant lymphoma, and that disease-related mortality would be much higher compared with radiation-induced mortality. This expectation is in accordance with a very recent publication by Brenner et al.   in adults –30. The 18F-FDG PET.Imaging with ultrasound and chest radiography were not considered in this study, as ultrasound is not associated with radiation exposure, and the radiation exposure from chest radiography is negligible compared with CT and There was no information available that allowed for assessment of organ doses in paediatric patients undergoing CT. Therefore we created Medical Internal Radiation Dose (MIRD) paediatric patient models in five general categories according to age and weight: newborn (3.6 kg), 1 year old (9.7 kg), 5 years old (19.8 kg), 10 years old (33.2 kg) and 15 years old (56.8 kg). The adult MIRD patient model represents an average-sized patient of 74 kg. All these hermaphrodite patient models (MIRD V) are described by spheres, ellipsoids and cones, as illustrated in Fig. In CT it is common practice to adjust acquisition parameters to the size of the patient, and to the clinical application . The acqDose calculations that were performed for the MIRD phantoms with the Monte Carlo software yielded organ doses, total body dose and effective dose according to ICRP 103. Dose assessment was performed for 12 CT acquisitions, i.e. both for whole-body CT , and for CT of the neck and chest only, in all six age categories. These doses were incorporated into the risk model, as described under “Risk assessment”. From these results, appropriate organ doses and the effective dose were derived for the year of diagnosis and treatment and for the following years of surveillance. If necessary, linear interpolation of dose values was performed to yield dose estimations at ages not included in the table.18F-FDG PET, published tables were used that provide information about organ dose and effective dose per MBq of administered 18F-FDG activity. The ICRP provides information for 1-, 5-, 10- and 15-year-old children, and for adults , resulted in an average 10-year survival of 94% (range 93–94%) [100% (range 99-100%)] for the young patient group with HD and an average 5-year survival of 55% (range 46–59%) [90% (range 76-98%)] for the adult patient group with DLBCL. Within the paediatric patient group, the average fraction of radiation-induced deaths was 0.4% [0.6%] for boys and 0.7% [1.1%] for girls. Within the adult group these values were even smaller; 0.07% [0.28%] for men and 0.09% [0.37%] for women. The average radiation-induced reduction of life expectancy in paediatric patients with HD is 21 days for boys and 45 days for girls. In adults with DLBCL, the average radiation-induced reduction of life expectancy is 1.5 days in men and 2.0 days in women.The calculations, which included disease-related mortality [or without disease-related mortality This study shows that the effect of disease-related mortality on radiation risk assessment in patients with malignant lymphoma is substantial. By taking into account disease-related mortality, the proportion of radiation-induced deaths decreased between 30% and 40% in paediatric patients with HD, and between 50% and 80% in adults with DLBCL. Radiation-induced reduction of life expectancy decreased by similar percentages. Our methodology, which integrates a model for organ dose assessment and risk assessment in one demographic model, showed that the potential radiation-induced reduction of life expectancy is only a small fraction compared with the disease-related reduction of life expectancy, namely a fraction between 0.003 and 0.011 for HD (children); and between 0.0003 and 0.0006 for DLBCL (adults). This reflects the fact that the radiation-related risk is a late risk with its expression up to decades after the actual exposure and has therefore less effect on the reduction of life expectancy. Our methodology for radiation risk assessment is an improvement compared with the usual standard of practice, where disease-related mortality is not considered. Although the observed cumulative effective doses are relatively high for imaging patients with malignant lymphoma, the associated estimated radiation risks are still very modest.Within the paediatric patient group, it is estimated that, on average, 0.4% of male and 0.7% of female patients eventually die because of the radiation exposure associated with medical imaging. For adult patients diagnosed with DLBCL, it is estimated that 0.07% of male and 0.09% of female patients die because of radiation exposure. The higher values for women result mainly from the relatively high sensitivity of the female breast to radiation exposure .Calculation in which the disease-related mortality was taken into account resulted in an average 10-year survival of 94%  for the young patient group with HD, which matches excellently with the published overall 10-year survival of 94% , 28. SimThis study has some limitations. Malignant lymphoma comprises a heterogeneous group of entities, with more than 40 subtypes . In addiFurthermore, with the implementation of integrated PET-CT systems, low-dose CT is performed in addition to PET, as it is used for attenuation correction in PET imaging. The use of CT for attenuation correction reduces the examination time, which implies a lower dose of FDG (5 MBq/kg vs 3 MBq/kg). The low-dose CT was not included in this study. Two or one additional low-dose CT(s) would have been performed in children with HD and in adults with DLBCL, respectively, with each low-dose CT causing extra radiation exposure of only approximately 3 mSv for adults . Finally18F-FDG PET-CT combined with a diagnostic CT, the most accurate and appropriate method of imaging during follow-up may therefore prove to be the low dose PET-CT, without the diagnostic CT. Another recent development is the introduction of whole-body magnetic resonance imaging (MRI) for the evaluation of malignant lymphoma. Initial results on this application of whole-body MRI are promising, but more research is still needed before it can be recommended as an alternative to CT and/or 18F-FDG PET [In order to establish the most efficient imaging strategy and limit the radiation exposure in patients with malignant lymphoma, there are different options. It may be sufficient to use only the low-dose whole-body CT (combined with FDG-PET) during and after therapy instead of a diagnostic (i.e. full-dose) whole body CT. This implies a reduction in radiation exposure of 6 times 8 mSv for adults. In patients with a FDG-avid type of malignant lymphoma and a baseline -FDG PET –46.18F-FDG PET is considered as justified in patients with malignant lymphoma, but should still be performed with care, especially in children. Ongoing studies have to establish the most efficient imaging strategies for (the different subtypes of) malignant lymphoma.In conclusion, the disease-related reduction in life expectancy of patients diagnosed with malignant lymphoma must be taken into account to achieve more realistic estimates of radiation risk. It results in higher overall mortality and substantial lower incidence of radiation-induced deaths. Although the cumulative effective dose from medical imaging is high, the actual calculated radiation risks are very modest. The radiation exposure that results from imaging with CT and


Epidemiological studies have found plasma lipid and lipoprotein levels to be predictive of cardiovascular disease in adults. To date, regular aerobic modes of exercise have been associated with favourable alterations in lipid and lipoprotein levels. However, the effect of resistance training on lipid and lipoprotein levels is inconclusive and conflicting. Therefore, the aim of this study was to provide some clarity on whether resistance training could be used to improve sedentary male smokers’ lipoprotein profiles.n = 13) or a control (CON) group (n = 12). Throughout the 16-week experimental period the CON group received no treatment whatsoever. After resistance training, serum triglyceride levels were significantly decreased by 18.42% from 1.162 mmol/l (± 0.476) to 0.831 mmol/l (± 0.058) (p = 0.038) in the RES group. However, resistance training was found to have no impact on any of the other measured lipid and lipoprotein measures.The study made use of a pre-test, a treatment period and a post-test. Subjects were placed into one of two groups, namely, a resistance-training (RES) group (In conclusion, these findings indicate that resistance training appears to have no significant effect on lipid and lipoprotein profiles in sedentary male smokers and therefore cannot prevent the advance of CAD. Atherosclerosis is the leading cause of coronary artery disease(CAD) in most Western populations and is associated with an accumulation of cholesterol in the muscular walls of arteries.3The terms ‘hyperlipidaemia’ and/or ‘hypercholesterolaemia’ should be considered misnomers since an increase in highdensity lipoprotein cholesterol (HDL-C) (hyperlipidaemia) must be seen as a negative risk factor and so the term dyslipidaemia should rather be utilised to describe this risk factor for CAD. Dyslipidaemia is generally characterised by hypertriglyceridaemia, an increased low-density lipoprotein cholesterol (LDL-C) and a decreased HDL-C.4Exercise training plays an important role in improving lipid and lipoprotein levels, since dietary or caloric restriction alone has been shown to be an ineffective method of improving lipid and lipoprotein profiles in the long term.9To date, regular aerobic modes of exercise have been associated with favourable alterations in lipid and lipoprotein levels.13Resistance training could be beneficial in improving lipid and lipoprotein levels since it results in an increased muscle lipoprotein lipase (LPL) and disparate decrease in hepatic LPL, which might result in favourable lipid and lipoprotein alterations. 14 The increase in muscle LPL and the decrease in hepatic LPL following resistance training may result in increased very low-density lipoprotein cholesterol catabolism and decreased HDL-C breakdown, respectively.14The aim of this study was therefore to investigate whether resistance training could improve a sedentary male smoker’s lipid and lipoprotein profile.n = 12) or resistance-training group (RES) (n = 13). Sedentary male smokers were selected as the target sample group since cigarette smoking and physical inactivity are two of the major CAD risk factors and both place the sedentary smoker at risk for developing an unfavourable lipid and lipoprotein profile.The study made use of a quantitative, pre-test/post-test research design. Twenty-five apparently healthy, sedentary male smokers between the ages of 20 and 35 years (mean age was 28 years and 7 months) volunteered to participate in the study. The subjects were informed about the potential risk of participation and gave their signed consent. Subjects were placed in either a control group (CON) , triglyceride (TG), LDL-C and HDL-C concentrations and TC:HDL-C, LDL-C: HDL-C and non-HDL-C (n-HDL-C) ratios/indexes were measured and/or calculated17The exercise intensity for each subject in the RES group was calculated subsequent to each subject undergoing a 10-repetition maximum evaluation for each prescribed exercise. The results of each subject were recorded in terms of the weight lifted and the amount of repetitions, which had to be less than 10, to reach fatigue. Each subject’s one-repetition maximum (1-RM) was then calculated according to the following formula: 1-RM = (weight lifted) ÷ [1.0278 − (repetitions to fatigue × 0.0278)].Members of the CON group were instructed to maintain their usual activities and not to take part in any form of structured exercise during the experimental period. As with the experimental group, they were also not given any advice on their diet, alcohol intake and smoking behaviour over the experimental period.The RES group subjects were instructed to exercise three times weekly for a period of 16 weeks. All exercise sessions were preceded by five minutes of easy cycling at a heart rate of less than 100 beats per minute (bpm), followed by eight stretching exercises performed for two sets of 30 seconds. Exercise sessions were concluded with five minutes of easy cycling at a heart rate of less than 100 bpm.13p ≤ 0.05) was used for determining a statistically significant change from the pre-test to the post-test.Descriptive statistics and dependant t-tests were calculated by the Rand Afrikaans University’s Statistical Consultation Service (STATKON) using the Statistical Package for Social Sciences (SPSS) II. A 95% confidence level (p = 1.000) p = 0.754). Similarly, the increase in HDL-C levels was found to be non-significant (p = 0.754). With regard to the calculated indexes, the n-HDL-C and TC:HDL-C ratio in the CON group decreased non-significantly by 0.780% (p = 0.530) and 0.260% (p = 0.638), respectively. The CON group’s LDL-C:HDL-C ratio increased non-significantly by 2.870% from the pre- to the post-test (p = 0.875). The CON group’s TG levels increased significantly  by 9.480% from 1.621 mmol/l to 1.761 mmol/l (p = 0.026).With regard to the CON group, a non-significant 0.350% decrease in TC levels was found at the conclusion of the 16-week experimental period (p = 0.807). Despite the RES group demonstrating a 7.000% increase in LDL-C levels, this change was found to be non-significant (p = 0.116). Similarly, the increase in HDL-C levels was found to be non-significant following RES training (from 1.052 mmol/l to 1.169 mmol/l) (p = 0.069) as was the 5.640% decrease in n-HDL-C levels (from 2.625 to 2.456) (p = 0.152). Although the RES group was found to have an 8.500% reduction in the TC:HDL-C ratio, this reduction was found to be non-significant (p = 0.173). Moreover, the 16 weeks of RES training resulted in a large mean  13.420% decrease in their LDL-C:HDL-C ratio (p = 0.152). In contrast, the RES training did result in a significant 18.420% decrease in TG levels from 1.162 mmol/l to 0.831 mmol/l (p = 0.038).Similar to the CON group, no significant change in mean TC concentration was observed following 16 weeks of RES training, with only a 1.570% increase being found at the post-test , is essential in determining CAD risk, few studies have focused on the effect of resistance training on this risk factor.The present study and other studies28The findings of the present study demonstrated that resistance training was not associated with favourable changes in lipid and lipoprotein levels in sedentary male smokers. Also, the sheer diversity of the different methods of applying resistance training is in itself a source of complication when attempting to determine if resistance training can indeed favourably alter lipid and lipoprotein profiles. Therefore, an optimal combination of resistance-training workload, intensity and number of repetitions still eludes researchers in this field. The contradictory findings in the literature, including the present findings, suggest that such an ‘optimal combination’ probably does not exist. It is therefore difficult to make a case for the use of resistance training to favourably alter the lipid and lipoprotein profiles of sedentary male smokers, and thus to reduce their risk of CAD.


Herein, we firstly demonstrate the design and the proof-of-concept use of a capillary-driven surface-enhanced Raman scattering (SERS)-based microfluidic chip for abrin detection. The micropillar array substrate was etched and coated with a gold film by microelectromechanical systems (MEMS) process to integrate into a lateral flow test strip. The detection of abrin solutions of various concentrations was performed by the as-prepared microfluidic chip. It was shown that the correlation between the abrin concentration and SERS signal was found to be linear within the range of 0.1 ng/mL to 1 μg/mL with a limit of detection of 0.1 ng/mL. Our microfluidic chip design enhanced the operability of SERS-based immunodiagnostic techniques, significantly reducing the complication and cost of preparation as compared to previous SERS-based works. Meanwhile, this design proved the superiority to conventional lateral flow test strips in respect of both sensitivity and quantitation and showed great potential in the diagnosis and treatment for abrin poisoning as well as on-site screening of abrin-spiked materials. Recently, ricin has caught the public's attention by the toxin-tainted letters sent to US President Barack Obama, Mississippi Senator Roger Wicker, and a Mississippi justice official, while abrin, its 70 times more toxic analogue, is less known to the general public. Abrin and ricin are toxic proteins with similar structure and properties, both of which are classified as category B select agents by the US Health and Human Services . Compare3 to 1015, which means that the technique may detect proper analytes at a single molecule level [Surface-enhanced Raman scattering (SERS) is a surface-sensitive technique that provides a highly enhanced Raman signal from Raman-active molecules that have been adsorbed onto rough metal surfaces. The reported surface enhancement factor ranges from 10le level . There ale level . Besidesle level . So far,le level -15. ThesHerein, we introduce a proof-of-concept use of the capillary-driven SERS-based microfluidic chip for abrin detection Figure . A microAll animal experiments (No. SYXK2007-0025) were approved by the Institutional Animal Care and Use Committee of Shanghai Jiao Tong University.Abrus precatorius (approximately 100 g) were soaked in 200 mL of 0.01 M phosphate buffer solution (PBS) at pH 7.4 and 4°C for 24 h. After thorough homogenization, the puree was centrifuged at 10,000g for 30 min. Then, the aqueous layer was saturated with ammonium sulfate (95% w/v) and centrifuged at 10,000g for 30 min. The precipitate was dissolved in 100 mL of 0.01 M PBS and applied to a 1.5 × 10 cm Gal-agarose column . The bound abrin was eluted with 0.125 M d-galactose solution. The collected fractions were dialyzed and applied to a Sephacryl S-100 prepacked column  equilibrated in PBS. The as-prepared abrin was analyzed by 15% sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE).Natural abrin was extracted according to the previous method with slight modifications . BrieflyThe purified abrin was inactivated by formalin and used to hyperimmunize a rabbit, and 0.5 mL of abrin toxoid (80 mg/mL) was mixed with an equal volume of Freund's complete adjuvant and injected subcutaneously to the rabbit. Seven days later, immunization was carried out four times including one booster immunization with the mixture of the abrin toxoid and Freund's incomplete adjuvant as well as three injections with the toxoid at weekly intervals. Ten days after the final injection, the immunized blood was collected by jugular puncture, and the serum was separated for subsequent purification of anti-abrin polyclonal antibodies with rProtein A Sepharose Fast Flow . The antibody titers were evaluated by enzyme-linked immunosorbent assay (ELISA).g for 1 h. Bovine serum albumin (BSA) was used to block the unmodified colloidal gold at a final concentration of 0.5% (w/v). The labeled colloidal gold was centrifuged at 12,000g for 1 h and resuspended in 1 mL 0.01 M PBS solution. Twenty microliters of DTNB solution (1 mM in 0.01 M PBS) was added to the gold solution and incubated at 4°C for 1 h. The resultant SERS probes were centrifuged again at 12,000g for 1 h and then resuspended in 0.01 M PBS for later use.The external SERS probes were prepared according to a published method . DTNB  using wet oxidation in a thermal furnace . Then, a photoresist  was spin-coated at 3,000 rpm to a thickness of approximately 20 μm and soft-baked for 90 min at 80°C. The layer was patterned subsequently by photolithography. The buffered hydrofluoric acid  was used to etch SiO2 uncovered by the photoresist. Afterwards, the microchips were etched into Si to a depth of 50 μm using deep reactive ion etching , and the micro silicon cylindrical array formed. The photoresist and SiO2 mask were removed by acetone  and DRIE, respectively. The as-prepared chips were cut into strips (1 × 4 mm) using a laser scribing apparatus . After being cleaned with the reactive ion etcher  at 30 W and 1.2 Torr for 45 s, the chips were then incubated in a solution of acetone for 20 min, rinsed with deionized water, and dried under an N2 stream. The deposition of gold film (200 nm) on the chip was carried out with the sputtering system . After being soaked in piranha solution (H2SO4/30% H2O2 = 3:1) for 10 min, the gold-coated chips were cleaned with deionized water and rinsed in 1 mM of HS-C2H4-CONH-PEG-C3H6-COOH  solution for 4 h. Finally, the chips were cleaned with deionized water and dried at room temperature. Scanning electron microscopy (SEM)  was used to explore the surface ultrastructure of the as-prepared chip. PBS was used to evaluate the flow rate of the sample solution on the chip.The gold-coated silicon wafer was fabricated by MEMS technique. The process was shown in Figure w/v). The SERS probes were printed on a glass fiber filter as conjugate pad and dried at room temperature. The absorbent pad, conjugate pad, and sample pad were cut into strips of 1 mm in width with a guillotine cutter and overlapped on the laminating card with the dried wafer as shown in Figure Anti-abrin polyclonal antibodies and goat anti-rabbit secondary antibodies 1 mg/mL) were dispensed on the gold-coated wafer with a Biodot XYZ3000 dispenser  as test zone and control zone, respectively. After drying for 30 min, the wafer was blocked with PBS containing 1% BSA  after 5 min. The intensity of the peak at approximately 1,330 cmA. precatorius was complicated due to the existence of abundant agglutinin that possesses nearly identical galactose-binding properties as abrin. Given their differences in galactose-binding avidity and molecular mass between the abrin and agglutinin, a two-step purification was exploited to separate abrin from raw extracts . Because of the absence of the washing step, some SERS probes remained on the gold-coated substrate, resulting in a weak nonspecific binding peak at approximate 1,330 cm-1. Figure y =1,430.7x - 2,312.5 and the correlation coefficient (R2) was 0.9902. The LOD of this capillary-driven SERS-based microfluidic chip was 0.1 ng/mL.Figure As previously mentioned, SERS-based techniques showed many potential advantages including high sensitivity, narrow bandwidths, and photobleaching resistance. It still remains a challenge to develop a SERS-based immunodiagnostic technique of both low cost and good operability. Some pioneering researchers have published their works focusing on the ultrasensitivity from the level of picograms per milliliter to femtograms per milliliter ,14,23-28We have designed and demonstrated the proof-of-concept use of our capillary-driven SERS-based microfluidic chip for abrin detection. The microfluidic chip was fabricated by MEMS process. The combination of the traditional LF test strip with capillary-driven gold-coated substrate results in the enhancement of sensitivity as well as the reduction of cost for SERS-based immunodiagnostic techniques. In this work, a calibration curve was obtained to detect the concentration of abrin in the range from 0.1 ng/mL to 1 μg/mL, which is superior to the traditional LF test strip for the same purpose in respect of both sensitivity and quantitation . What isThe authors have declared that no competing interest exists.HY carried out antibody preparation and SERS experiments. MD finished the microfabrication of the micropillary chip. SG finished the surface modification of the micropillary chip. SHC finished the antibody conjugation with the surface of the chip. LK and JW finished the characterization of the chip. WX, TZ, and ZY finished the result analysis. HY and YA finished the draft. JW and DC finished the experiment design and manuscript revision. All authors of this paper have read and approved the final manuscript.


The major radiation of the Sun can be roughly divided into three regions: ultraviolet, visible, and infrared light. Detection in these three regions is important to human beings. The metal-insulator-semiconductor photodetector, with a simpler process than the pn-junction photodetector and a lower dark current than the MSM photodetector, has been developed for light detection in these three regions. Ideal UV photodetectors with high UV-to-visible rejection ratio could be demonstrated with III–V metal-insulator-semiconductor UV photodetectors. The visible-light detection and near-infrared optical communications have been implemented with Si and Ge metal-insulator-semiconductor photodetectors. For mid- and long-wavelength infrared detection, metal-insulator-semiconductor SiGe/Si quantum dot infrared photodetectors have been developed, and the detection spectrum covers atmospheric transmission windows. UV light is usually divided into three regions: UV-A, UV-B, and UV-C . UV-C , and metal-insulator-semiconductor (MIS) structures ,5. When This review focuses on MIS photodetectors working in the UV, visible or infrared regions. First, the electrical characteristics of the MIS structure will be discussed. The MIS UV photodetectors using wide-bandgap semiconductor materials or metal filters will be discussed in Section 3. Then, the MIS structures for visible-to-near-IR detection employing the most broadly used semiconductor material, Si, will be discussed. Ge can be used to extend the cutoff wavelength of Si photodetectors. Finally, mid- and long-wavelength infrared detection by HgCdTe MIS photodetectors and MIS SiGe/Si quantum dot (well) infrared photodetectors will be introduced in Section 5.2.MOSFETs have become basic components of most electronic devices. In order to increase circuit performance, the quest to scale down MOSFETs continues. As the device size is scaled down, the oxide thickness decreases and the gate tunneling current along the vertical MOS structure  then increases. 2/p-Si diode for light detection. From .,52.45,52Due to the Si bandgap, the near infrared at the optical communication wavelengths of 1,300 and 1,550 nm cannot be detected. The detection performance at the wavelength of 820 nm is also limited. Five periods of SiGe quantum-dot layers were incorporated into the Si substrate to improve the near-infrared performance of MIS photodetectors . The SiG4.2.2 layer was formed on the Ge substrate with the process of low-temperature liquid phase deposition (LPD) et al. [7et al. showed that the fixed charge density of 3.2 × 1011 cm−2 of the sulfide treated MIS sample was smaller than that of 6.4 × 1011 cm−2 of the untreated sample in [11 cm−2 to 7.5 × 1010 cm−2 after sulfidation. Such improvement was due to the suppression of native oxidation on the surface with the sulfidation process.Jung ample in . The slo2O3, and SiO2 layers. The native oxide layers were formed on an insulated holder outside the plasma, and the ion bombarding was avoided. The fixed charge density and interface trap density were as low as 1.5 × 1011 cm−2 and 3 × 1011 cm−2 eV−1. respectively. Al2O3 and SiO2 layers were formed by RF magnetron sputtering with 0.011 Pa of oxygen in an argon atmosphere. Their fixed charge densities and interface trap densities were smaller than 6.0 × 1010 cm−2 and 5 × 1011 cm−2 eV−1. The interface quality is high enough to eliminate the Fermi level pinning. It could be expected that continuous investigation on HgCdTe photodetectors based on the MIS structure will be performed. However, it will be a difficult assignment for the HgCdTe MIS photodetectors to compete with the HgCdTe pn-junction-type photodetectors.Different passivation insulator layers were also used to suppress Fermi level pinning of HgCdTe MIS photodetectors . These i5.2.Due to the natural valence band offset at the SiGe/Si heterojunction, discrete energy levels could be formed inside the Si/SiGe/Si quantum confinement region. The transition between these discrete levels can be used for infrared detection. The SiGe/Si quantum dots and quantum wells were broadly investigated for mid-infrared and long-wavelength detection –76. The It is worth mentioning that the suitable gate electrode selected here is Al instead of Pt, which has been suggested in The J-V curves of SiGe/Si QDIPs with and without the insertion of an insulator layer between the metal and semiconductor are compared in 0.9Ge0.1 quantum wells. For the spacer sample, the detection regions of 3.7–6 and 6–16 μm corresponded to the LH1-to-LH3 transition in SiGe/Si QDs, and intraband transitions in boron doping wells, respectively.Spectral responses of MIS SiGe/Si quantum dot (well) infrared photodetectors are shown in 0.9Ge0.1 quantum wells would be very shallow unlike that of SiGe QDs , the LH of ∼0.5 .6.Nowadays, integrated circuits are composed of MOSFETs. If the MIS structure can be utilized for photodetectors, the integration of detection devices and readout circuit will be much easier and less expensive to manufacture. The wide-bandgap semiconductor has been utilized in the MIS UV photodetectors, and the optimized UV-to-visible rejection ratio could be more than three orders of magnitude. Si-based and thin-film Ge MIS photodetectors are suitable for visible-to-near-IR photo-detection. With the MIS SiGe/Si QDIPs, a broadband infrared detection covering atmospheric transmission windows at 3–5.3 μm and 7.5–14 μm is possible.


Cognitive function was evaluated by two behavioral tests  in early (4–6 months of age) and late phase (7–12 months of age). Serum MG was significantly elevated in the MG group  at the end of study. The groups did not differ in cognitive function during the course of study. No time-course differences were found in oxidative stress markers between the two groups, while, antioxidants such as glutathione peroxidase and superoxide dismutase activities were significantly increased in the MG group compared to the control. Long-term MG administration to rats with normal kidney function did not cause CI. A counter-balanced activation of the systemic anti-oxidant system may offset the toxicity of MG in this model. Pathogenetic significance of MG for CI requires further investigation.Methylglyoxal (MG), one of the uremic toxins, is a highly reactive alpha-dicarbonyl compound. Recent clinical studies have demonstrated the close associations of cognitive impairment (CI) with plasma MG levels and presence of kidney dysfunction. Therefore, the present study aims to examine whether MG is a direct causative substance for CI development. Eight-week-old male Sprague-Dawley (SD) rats were divided into two groups: control ( Methylglyoxal (MG) is a highly reactive alpha-dicarbonyl and protein-bound type compound with a molecular weight of 72 dalton . MG bindet al. reported that AGEs in cerebrospinal fluid were associated with CI [Accumulative clinical evidence has revealed that cognitive impairment (CI) is commonly prevalent among patients with a various stages of chronic kidney disease (CKD), ranging from early to advanced stage, including cases on dialysis treatment ,4,5,6,7. with CI . MG is o with CI . Taken tIt is still unknown whether MG is a direct causative substance of CI, including whether it plays a role in the pathology and mechanism of CI. Although previous researchers have demonstrated MG-induced cytotoxicity in cultured hippocampal neurons and cerebral cortex neuronal cells ,13, it iThe aim of the present study is to confirm whether long-term orally administered MG induces CI in Sprague-Dawley (SD) rats with normal kidney function. vs. control, 495.8 ± 38.1 vs. 244.8 ± 28.2 nM (p < 0.001)  . The driThe results of object exploration tests in both phases are shown in p = 0.001) and in Session 11–15 (p = 0.017). The number of working memory errors  during each Phase is shown in We examined the radial-arm maze test between ages of 20 and 23 weeks (Phase I), and between the ages of 42 and 44 weeks (Phase II). The results of the maze test, which was conducted to examine the spatial reference memory and spatial working memory, are shown in The central nervous system (CNS) shows vulnerability to toxic compounds such as MG. Accumulation of MG in the hippocampus has an important role in learning impairment and memory disorders . CA1 andvs. the control, 5.1 ± 0.4 vs. 3.6 ± 0.4 mU/mg (p = 0.036)). Superoxide dismutase (SOD) activity tended to be higher in the MG-treated group ). Urinary and plasma angiotensinogen (AGT) levels were significantly lower in the MG-treated group: MG vs. control, 2.1 ± 0.2 vs. 3.2 ± 0.3 ng/mL (p = 0.014), 16.3 ± 6.7 vs. 29.3 ± 6.9 ng/day (p = 0.034), respectively. These results are shown in We evaluated biomarkers using urine, blood and kidney samples . UrinaryThe present study aims to examine whether MG is a direct causative substance for CI development.ad lib drinking water for up to 12 months. We examined the cognitive function using two behavioral tests  in an early (4–6 months of age) and a late phase (7–12 months of age). Although serum MG was significantly elevated in the MG group, the groups did not differ in cognitive function during the course of study, and no histological changes were observed at the end of study. No time-course differences were found for the oxidative stress marker, while antioxidants were significantly increased in the MG group compared to the control.Eight-week-old male SD rats were given MG by N-carboxyethyllysine) is thought to be useful.Before we discuss our speculations of the results, we need to describe some significant limitations of our study. (1) We selected SD rats with normal kidney function; and (2) we could not confirm the MG status in the CNS. As to the first limitation, a kidney injury model might represent very different responses to long-term low doses of oral administered MG, because the interaction of MG with other uremic toxins or with oxidative stress is anticipated in those models. It might also be possible to detect the pathogenesis of the relationship between CI and MG by using diabetes-model animals. As to the second limitation, we failed to examine the MG status in the CNS. The serum MG level was significantly increased in the MG group rats, however, the impact of MG on the CNS was unclear, because we could not detect how much MG passed through the blood brain barrier and reached the CNS. This suggests that a significant amount of MG is thought to be needed in order to enter the brain from the blood. Additionally, in terms of evaluation of the MG status in the CNS, measuring brain and kidney MG levels or MG-induced AGEs , blood pressure elevation, and diabetic-like pathological changes ,17,18. ICognitive impairment was not demonstrated by MG loading in this study. One of the acceptable reasons for our results is the MG concentration. The plasma MG levels were thought to be relatively low to induce neuronal damage. Demonstrating the CI based on the orally available dose of MG may be difficult. A high concentration of MG  has been employed in previous studies using cultured hippocampal neurons ,26. Addiet al reported that MG reacted characteristically with hydrogen peroxide, and that methyl- and carbon-centered radicals were generated non-enzymatically [Nevertheless, it is not possible to exclude the possibility of MG as a candidate substance for CI based only on our limited data. MG is an uremic toxin, which induces and/or amplifies oxidative stress ,29,30. Natically . ConsideThe activation of the antioxidant system observed in our protocol is interesting. We speculate that a counter regulation against MG exposure up-regulated the antioxidant system. This phenomenon is sometimes described as “hormesis effect”. Hormesis is the phenomenon by which an organism shows a favorable biological response to low-dose exposures to stressors such as toxic compounds and radiation . The horYoung adult male SD rats at 8 weeks of age  were housed under controlled environmental conditions  with free access to water and standard pellet food. All procedures were conducted in accordance with the National Institutes of Health Guide for the Care and Use of Laboratory Animals, and the study protocols were approved by the animal committee of Fukushima Medical University . n = 9; tap drinking water), and the MG group  as drinking water). The rats were fed with standard pellet food . Body weight was measured at 8, 16, 23, 31 and 47 weeks of age, and blood pressure and heart rate were measured at 8, 16, 20 and 47 weeks of age by an indirect tail-cuff method . Timeline chart of the experiments and physical findings are shown in The rats were divided into two groups: the control group  object exploration test for assessing spatial memory; and (2) eight-arm radial maze test for assessing spatial memory-related reference and working memory.Object exploration test was examined at 16 (Phase I) and 31 weeks of age (Phase II). The device and situation of the test are shown in Next, we examined the spatial working and reference memory using the radial-arm maze test based on the protocol reported previously . The weiTwenty-four-hour urinary samples were collected from the rats in a metabolic cage at the ages of 16 (Phase I) and 45 weeks (Phase II). All animals were sacrificed after the behavior tests were completed at the age 47 weeks. Pentobarbital (50 mg/kg) was administered intraperitoneally for euthanasia. Then the abdominal cavity was opened and as much blood as possible was collected from the descending aorta. Next, saline was perfused. After blood removal was completed, the bilateral kidneys were removed. Then the thorax was opened and perfused transcardially with phosphate-buffered saline (PBS) followed by 4% paraformaldehyde in 0.1 M phosphate buffer of pH 7.4. Methylglyoxal was measured using liquid chromatography/mass spectrometry, as previously reported .in vitro, which reflects the degree of oxidative stress [et al. [et al. [In Phase I, we examined urinary 8-hydroxy-2'-deoxyguanosine (8-OHdG) and malondialdehyde (MDA). In addition to these two oxidative stress markers, we examined glutathione peroxidase (GPx) activity, superoxide dismutase (SOD) activity in the kidneys as an antioxidant marker, and angiotensinogen (AGT) in the urine and plasma were also examined in Phase II by a commercially available kit. We asked a laboratory testing company  to measure uric protein, uric and serum creatinine, blood urea nitrogen, plasma glucose and urinary 8-OHdG to. Urinary 8-OHdG is a marker of damage by DNA oxidation e stress . MDA hase stress . We meas [et al. . GPx act [et al. . Enzyme  [et al. . The SOD [et al.   staining and caspase-3 staining. Fixed brains were cut into sections (6 µm thick) through the sagittal plane with a cryostat for H&E and caspase-3 staining. For caspase-3 staining, a rabbit anti-cleaved caspase-3 (Asp175) antibody  was used. Rat kidney histological examination was conducted under light microscopy with periodic acid-Schiff staining. Kidney samples obtained from each rat were fixed in 10% buffered formalin and embedded in paraffin and serially sectioned at 2.5 µm thickness. post hoc test was used. Data analyses of the physical findings and laboratory tests, including serum MG and oxidative-stress or antioxidative markers and angiotensinogen (AGT), were performed using Student’s t test. Values of p < 0.05 were considered to indicate statistical significance. Statistical analyses were performed using IBM SPSS Statistics version 19 . All results were expressed as the mean ± standard error of the mean. Comparisons of means of the groups of the two behavior tests were analyzed by repeated one-way analysis of variance (ANOVA). After ANOVA, Turkey-Kramer’s in vitro. The significance of MG in the pathogenesis of CI requires further investigation. Long-term oral administration of MG in SD rats with normal kidney function did not cause CI, despite a significant increase in serum MG at the level of advanced CKD. Counter-balanced activation of the systemic anti-oxidant system by the MG load may offset the toxicity of MG in this model. Our results are considered to be important fundamental data for investigating MG toxicity under various conditions


Background: The underlying molecular mechanisms of PTSD are largely unknown. Distinct expression signatures for PTSD have been found, in particular for immune activation transcripts. DNA methylation may be significant in the pathophysiology of PTSD, since the process is intrinsically linked to gene expression. We evaluated temporal changes in DNA methylation in select promoter regions of immune system-related genes in U.S. military service members with a PTSD diagnosis, pre- and post-diagnosis, and in controls.Methods: Cases (n = 75) had a post-deployment diagnosis of PTSD in their medical record. Controls (n = 75) were randomly selected service members with no PTSD diagnosis. DNA was extracted from pre- and post-deployment sera. DNA methylation (%5-mC) was quantified at specific CpG sites in promoter regions of insulin-like growth factor 2 (IGF2), long non-coding RNA transcript H19, interleukin-8 (IL8), IL16, and IL18 via pyrosequencing. We used multivariate analysis of variance and generalized linear models to calculate adjusted means  to make temporal comparisons of %5-mC for cases (pre- to post-deployment) versus controls (pre- to post-deployment).Results: There were significant differences in the change of %5-mC pre- to post-deployment between cases and controls for H19  and IL18 . For H19 the difference was driven by a significant reduction in %5-mC among controls; for IL18 the difference was driven by both a reduction in %5-mC among controls and an increase in %5-mC among cases. Stratified analyses revealed more pronounced differences in the adjusted means of pre-post H19 and IL18 methylation differences for cases versus controls among older service members, males, service members of white race, and those with shorter deployments (6–12 months).Conclusion: In the study of deployed personnel, those who did not develop PTSD had reduced %5-mC levels of H19 and IL18 after deployment, while those who did develop PTSD had increased levels of IL18. Additionally, pre-deployment the people who later became cases had lower levels of IL18 %5-mC compared with controls. These findings are preliminary and should be investigated in larger studies. Estimates of post-deployment PTSD prevalence in U.S. military service members who served in Afghanistan and Iraq are 12 and 18%, respectively , IL8, IL16, and IL18  had existing pre- and post-deployment serum samples housed at the DoDSR, met all the criteria above, and had at least two outpatient records with a primary diagnosis of chronic PTSD, based on ICD-9 Code 309.81 in the first diagnostic position. The first outpatient diagnosis was between 4 and 12 months after return from first deployment. The second outpatient diagnosis was any time after that, but within 2 years of return from first deployment. Additional criteria for inclusion as a case to this study was having one serum sample (the pre-deployment sample) drawn within 12 months prior to first OEF/OIF deployment and one sample (the post-deployment sample) drawn within 6 months after return from first OEF/OIF deployment. Cases were randomly selected within each gender, such that 25 females and 50 males were included. We identified an appropriate active duty service member control group (n = 75), who were frequency matched from a stratified, random sample based on age , gender, and race, but for whom there was never a diagnosis of PTSD (ICD-9 Code = 309.81) or Traumatic Brain Injury (TBI) . Though cases in this study are presumed to have experienced a PTE by virtue of their PTSD diagnosis, the criteria of which requires exposure to a PTE, we have no data on formal assessment of trauma/PTE for either cases or controls. As such, this study uses deployment as a proxy for exposure to a PTE for both cases and controls. Because of Department of Defense policy regarding the DoDSR samples, we were unable to make contact with the study subjects for whom we had serum and medical encounter data.The PTSD cases  and quantified via Qubit ®  dsDNA HS Assay Kit using a Qubit fluorometer .For each PTSD case and control, The Armed Forces Health Surveillance Center identified a pre-deployment and a post-deployment serum sample and authorized release of up to 0.5 ml of serum per sample. These samples had been maintained at −30 °C continuously at the DoDSR. DNA was extracted using ChargeSwitch DNA methylation was quantified via bisulfite treatment, PCR, and pyrosequencing. DNA was bisulfite treated using the Zymo DNA Methylation Kit . All assays used in this study were validated by PCR bias testing and sensitivity testing. This was done to assess the lowest input gDNA needed for bisulfite modification and the lowest input bisulfite converted DNA for PCR for each assay. For H19, IL8, IL16, and IL18, a minimum of 40 ng of DNA was used for bisulfite conversion. For IGF2, a minimum of 10 ng DNA was used for bisulfite conversion. The total DNA available for each sample was used as input gDNA for bisulfite conversion. Carrier RNA from Qiagen was used during the bisulfite modification and purification to minimize the loss of DNA during the procedure. Bisulfite treated DNA was eluted in 20 μl volume and 1 μl of it was used for each PCR. The PCR (45 cycles) was performed with one of the PCR primers biotinylated to convert the PCR product to single-stranded DNA templates. The PCR products (each 10 μl) were sequenced by Pyrosequencing PSQ96 HS System (Qiagen Pyrosequencing) following the manufacturer’s instructions (Qiagen Pyrosequencing). Pyrosequencing is a real-time sequencing-based on mutation analysis or methylation analysis technology. The methylation status of each CpG locus was analyzed individually as a T/C SNP using QCpG software (Qiagen Pyrosequencing).The loci of specific CpGs measured in each promoter region are shown in Figure p < 0.01), IL8 , IL16 , and IL18 . The duplicate samples were run on a separate plate from their counterparts, and some of the variation could be due to a plate effect.DNA from cases and controls were randomly arranged on the plates for methylation quantification analysis. For 10% of the samples we included duplicates to which laboratory personnel were blinded, for the purpose of quality control (QC). Coefficients of variation as well as intraclass correlation coefficients (ICCs) were calculated for the means of each position in H19  had DNA yields  insufficient for bisulfite conversion of H19, IL8, IL16, and IL18. Approximately 5% of the samples (N = 15) had DNA yields  insufficient for bisulfite conversion of IGF2. These samples were excluded from our study. For a small percentage of the samples for which bisulfite conversion was complete, pyrosequencing signals were too low to be reliable , thus we did not include them in our final analyses. It is unclear why signals were low for those particular samples, however, low signals were not associated with DNA yield. Distribution of samples either not measured or measured but with low signals did not differ by pre- and post-deployment or case/control status, with the exception of IGF2 pre-deployment case/control status. However, this is likely because there were only 16 with low signals for that assay, thus lending to the instability of those proportions.Approximately 30% of the samples  measured as well as the mean of all the positions measured. We compared the change in cases’ %5-mC from pre- to post-deployment, to the change in controls’ %5-mC from pre- to post-deployment using multivariate ANOVA. We present the adjusted mean of pre-post methylation difference for cases versus controls, adjusted for age, gender, and race. To further investigate the adjusted means and differences between cases and controls, pre-deployment and post-deployment, we ran generalized linear models (GLMs) Using GLMs, we carried out case-case and control-control  comparisons, as well as case-control comparisons, pre-deployment, and post-deployment. We carried out the same analyses, stratifying by age, gender, race, and length of deployment, separately, to investigate any potential patterns within specific groups. SAS Version 9.2 was used to carry out all statistical analyses. The SAS procedures GLM and ANOVA were used to run our final models  and IL18 . Position 3 in H19 and positions 4 and 5 in IL18 had statistically significant differences between cases and controls, and though differences in the change of methylation were not significant at other positions for H19 and IL18, they all showed similar patterns.Table Table S1 in Supplementary Material provides additional detail on the change in methylation across deployment for cases and controls; paired differences in adjusted mean levels for cases pre- and post-deployment and controls pre- and post-deployment, determined via GLM, are presented. The differences between cases and controls across deployment for H19 were driven primarily by the significant reduction in %5-mC among controls from pre- to post- deployment (the levels for cases were unchanged across deployment for H19). The pattern of decreased H19 methylation among controls from pre- to post-deployment is evident; statistically significant decreases were found at positions 1, 3, and 4, and for the mean of positions 1–4. The differences for IL18 were driven by both the decrease in methylation across deployment for controls as well as the increase in methylation across deployment for cases. There was a pattern of decreased methylation among controls from pre- to post-deployment, though a statistically significant decrease was found for position 5 only. There was also a pattern of increased IL18 methylation for cases pre- to post-deployment, though none of the comparisons was statistically significant.p ≤ 0.05 for positions 2, 4, and 5, and the mean of the five positions), while the pattern reversed post-deployment, such that levels were non-statistically significantly higher in cases compared with controls. Note that the term “case” in the pre-deployment context refers to people who later became cases, post-deployment; during the pre-deployment time period, they were not yet cases.Table S2 in Supplementary Material, which presents case-control comparisons for both pre- and post-deployment, further illustrates that for IL18, %5-mC was lower in cases compared with controls, pre-deployment  and IL8 position 2 . There were no significant differences in adjusted mean levels for the post-deployment case-control comparisons.Also shown in Table S2 in Supplementary Material are a few other loci where significant differences in %5-mC were found between cases and controls pre-deployment, i.e., IGF2 position 2 (%5-mCp = 0.05), males , service members of white race , and those with deployments between 6 and 12 months .Stratified analyses Table  revealedOur study found that there were significant differences in the change of methylation (%5-mC) across deployment between cases and controls for H19 and IL18. While there was no change in %5-mC in H19 for cases from pre- to post-deployment, there was a significant decrease among controls driving the difference in %5-mC change between the cases and controls. For IL18 there was a significant difference between the %5-mC decrease in controls and the %5-mC increase in cases, from pre- to post-deployment. These findings were more pronounced in both H19 and IL18 in younger service members, males, service members of white race, and those with shorter deployment length. There were a few additional statistically significant findings at specific CpG sites, such as pre-deployment case-control differences in IGF2 position 2 and in IL8 position 2. Given the low signals from the IGF2 assay being differentially distributed between pre-deployment cases and controls, we are cautious to interpret the results for IGF2. Whether these patterns represent markers of vulnerability or resiliency is speculative. Psychological stress incurred during deployment may be associated with a methylation response in these genes, but given the lack of PTE data for controls, this study is not able to address whether the response is protective of PTSD or puts one at risk for PTSD.p-values approached significance for positions 1 (0.08) and 3 (0.06). There was a reversal of this pattern post-deployment, in that cases had higher levels at all positions, compared to controls, though differences were not statistically significant. Given that DNA methylation is often inversely correlated with gene expression, in particular for immune system-related genes . The imprinted IGF2-H19 locus (CTCF binding cite) encodes the growth promoting hormone IGF2 and a long non-coding RNA H19. The maternally inherited locus transcribes untranslated H19 RNA, which plays the role of a trans-regulator of the imprinted gene network controlling embryonic growth and was recently shown to be processed into micro-RNA (miRNA)-675 -receptor subunits, and neuronal phosphatases through its PDZ domain and exerts its autocrine function in the brain with a distinct role in neuropoiesis . The mean storage time for cases post-deployment was 3.3 years, for controls it was 3.7 years (p = 0.14). As we expected, the storage time did differ by pre- and post-deployment time period. Thus for cases pre-deployment the mean was 4.5 years, while post-deployment it was 3.4 years (p = < 0.01). Likewise, the mean storage time for controls pre-deployment was 4.9 years, while post-deployment it was 3.7 (p < 0.01). Based on these differences, we adjusted all our models in which we carried out a pre/post comparison   are directly proportional to the number of molecules being sequenced. Pyrosequencing QCpG software has a number of quality assessments to pass or to fail results for each sample being sequenced. These specifications give information including sample quality, non-specific PCR amplification, non-specific sequencing, and completion of bisulfite modification. Therefore, if a pyrosequencing run on a specific sample gave a passing result, it indicates that the DNA of that sample is of sufficient quality.Chance could account for some of the statistically significant findings we report, given the number of comparisons and the small sample size, but this was an exploratory study and the results should be considered as preliminary. The DoDSR, which contains over 45 million longitudinally collected serum samples of U.S. military service members will provide a vast resource for carrying out future investigations in larger populations. There is growing evidence that the molecular mechanisms that regulate DNA methylation are involved in synaptic plasticity, learning, and memory (Sananbenesi and Fischer, We evaluated DNA methylation from serum as a potential biomarker for PTSD. In this exploratory study of deployed personnel, comparing those with PTSD and those who did not develop PTSD, the latter had reduced %5-mC levels of H19 and IL18 after deployment and elevated levels of IL18 after deployment. Additionally, pre-deployment the people who later became cases had lower levels of IL18 %5-mC compared with controls. Whether these are markers of vulnerability or resilience is a matter of speculation, but because of the high incidence of PTSD, it is important to study these patterns further.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.http://www.frontiersin.org/Molecular_Psychiatry/10.3389/fpsyt.2013.00056/abstractThe Supplementary Material for this article can be found online at Supplementary Table S1∗ and differences between controls pre- and post-deployment and cases pre- and post-deploymentGeneralized linear models for investigating the adjusted means.Click here for additional data file.Supplementary Table S2∗ and differences between cases and controls, pre-deployment and post-deploymentGeneralized linear models investigating the adjusted means.Click here for additional data file.


Anti-TNF drugs have proven to be effective against spondyloarthritis (SpA), although 30% of patients fail to respond or experience adverse events leading to treatment discontinuation. In rheumatoid arthritis, the presence of anti-drug antibodies (ADA) against the first TNF inhibitor influences the outcome after switching. Our aim was to assess whether the response to a second anti-TNF drug is related to the previous development of ADA to the first anti-TNF drug SpA patients.Forty-two SpA patients began a second anti-TNF drug after failing to respond to the first anti-TNF therapy. Clinical activity was assessed by the Ankylosing Spondylitis Disease Activity Score (ASDAS) at baseline (at the beginning of the first and second anti-TNF therapy) and at 6 months after switching. The drug and ADA levels were measured by ELISA before each administration.p = 0.399) and to the second anti-TNF drug . At 6 months after switching, patients with previous ADA had lower disease activity  and most patients without ADA had high disease activity state by the ASDAS (25 out of 31 (80.6%) without ADA vs. 3 out of 11 (27.3%) with ADA, p = 0.002).All patients were treated with anti-TNF drugs and mainly due to inefficacy were switched to a second anti-TNF drug. Eleven of 42 (26.2%) developed ADA during the first biologic treatment. At baseline, no differences in ASDAS were found in patients with or without ADA to the first anti-TNF drug (3.52 ± 1.03 without ADA vs. 3.14 ± 0.95 with ADA, In SpA the failure to respond to the first anti-TNF drug due to the presence of ADA predicts a better clinical response to a second anti-TNF drug. Spondyloarthritis (SpA) describes a group of diseases including ankylosing spondylitis (AS), psoriatic SpA, SpA related to inflammatory bowel disease (IBD), reactive arthritis, a subgroup of juvenile idiopathic arthritis and nonradiographic axial spondyloarthritis . SeveralThe available anti-TNF drugs differ in chemical structure, half-life, route of application and capacity to induce immunogenicity, and they also have somewhat different mechanisms of action ,11. AlthTo date, only two studies have been published that correlate the clinical response and immunogenicity to anti-TNF drugs in rheumatoid arthritis (RA) patients who switched to a second anti-TNF drug ,22. In tA total of 42 SpA patients  without previous biological treatment were included. All of these patients had axial involvement and most of them had some peripheral articular manifestation as dactylitis, enthesopathy, monoarthritis and oligoarthritis (28/42 (66.7%) SpA patients: 13 AS, 10 nonradiographic axial SpA, 2 psoriatic SpA, 2 SpA related to IBD and 1 reactive arthritis. The patients were enrolled at the Department of Rheumatology of La Paz University Hospital. This was an ambispective observational study that was approved by the La Paz Hospital Ethics Committee, and all patients provided informed written consent. The retrospective study period covered the years 2005 to 2008, and the prospective study period covered 2009 to 2011. All of the AS patients fulfilled the New York revised criteria for AS . The psoAll patients received anti-TNF drugs as a first biological treatment (infliximab (Ifx), adalimumab (Ada) and etanercept (Etn)) and later switched to a second anti-TNF drug ). The selection of all anti-TNF drugs was left to the discretion of the physician, with consideration of patient characteristics, type of disease, and patient preference. Owing to the observational design of the study, no specific criteria for drug withdrawal were required, and the diagnoses of treatment failure and adverse events were based on the judgement of the treating physician. Ifx was administered intravenously at 5 mg/kg at 0, 2 and 6 weeks and every 8 weeks thereafter, and the remaining anti-TNF drugs were administrated subcutaneously .Disease activity was measured by the Ankylosing Spondylitis Disease Activity Score (ASDAS) ,26 and wBlood samples were collected a maximum of 24 hours before biological drug administration for subcutaneous anti-TNF or just before intravenous infusion for Ifx. Precise timing was required to compare the results because the drug levels in the serum can become undetectable over longer time intervals as a result of normal drug pharmacokinetics rather than the formation of immunocomplexes with ADA. All sera, including those of the retrospective period, were stored at -20°C until the drug and ADA concentrations were measured.The serum drug concentrations  were determined by sandwich ELISA, as described previously -29. SeruSerum ADA levels  were detected using a two-site (bridging) ELISA, as previously described -29. The To determine the cutoff value of each assay, sera from 150 healthy controls and from 100 RA patients without anti-TNF treatment (70% positive for rheumatoid factor) were studied. The mean ± 6 standard deviations was used to establish cutoff points.The statistical analyses were performed using the Statistical Package for the Social Sciences, version 11.0 . Descriptive statistics included the mean and standard deviation or the median and interquartile range. Differences in baseline characteristics were assessed using Pearson's chi-square test and Fisher's exact test for ordinal variables and using the Mann-Whitney U test for continuous variables. The continuous data were compared between groups using the Mann-Whitney U test. Statistical significance was calculated using the log-rank test, and p < 0.05 was considered significant.A total of 42 SpA patients were enrolled in this study with a mean ± standard deviation age of 49.6 ± 10.4 years at the time of inclusion, and 23 (54.8%) were men. The baseline demographic and clinical characteristics of the global patient population, categorised according to future ADA development against the first anti-TNF therapy, are shown in Table All 42 patients were treated with an anti-TNF drug as the first biologic therapy  and were switched to a second anti-TNF treatment  due to inefficacy  and/or adverse events . Of the eight patients who withdrew due to adverse events, three had been treated with Ifx  and five with Etn .p = 0.006). ADA appeared mainly within the first year of anti-TNF therapy (mean ± standard deviation: 12.89 ± 5.92 months), except for five patients in whom ADA were detected at 18 months (2 patients), 20 months (2 patients) and 28 months (1 patient). At 6 months after switching to the second anti-TNF drug, ADA were detected in only two patients treated with Ifx, who had not previously developed antibodies against the first anti-TNF drug. The drug and ADA concentrations were not evaluated for Gol.ADA were detected in 11 (26.2%) patients (7/27 (25.9%) AS, 3/10 (30%) undifferentiated SpA and 1/2 (50%) SpA related to IBD) during treatment with the first anti-TNF drug and were more frequent in patients treated with Ifx (9 (81.8%) with Ifx, 2 (18.2%) with Ada, 0 (0%) with Etn, Most patients without ADA (28/31) against the first anti-TNF drug had clinical inefficacy and detectable serum drug levels just before the switch to the second anti-TNF drug (median (interquartile range): 3,008  ng/ml for Ifx, 3,072  ng/ml for Ada, 1,111  ng/ml for Etn), so these patients were considered as having a primary inefficacy. Only three patients treated with Etn without detectable antibodies before switching did not have loss of efficacy and they were switched due only to adverse events. Furthermore, all of 11 patients with ADA against the first anti-TNF treatment had loss of efficacy and undetectable drug levels before switching, so they were classified as having secondary inefficacy related to development of immunogenicity.p = 0.399; baseline ASDAS second anti-TNF: 3.36 ± 0.94 without ADA vs. 3.09 ± 0.91 with ADA, p = 0.466). Also, there were no differences in clinical activity at baseline to the first and second anti-TNF drugs in patients with and without ADA .At baseline, no differences in disease activity were observed between patients who did or did not later develop ADA against the first anti-TNF drug     . Three out of the five patients without ADA who had an important clinical improvement after switching were treated with Etn as the first anti-TNF drug and the reason for change to a second anti-TNF drug was adverse effects.At 6 months after switching we observed a greater clinical improvement, as measured by change in ASDAS, in patients with ADA as compared with those without ADA  Figure . A totaln = 27), we observed that our results in relation to clinical activity and immunogenicity are consistent with those observed analysing all 42 patients. At baseline of the first and second anti-TNF drugs, no differences were seen in clinical activity (ASDAS) between patients with and without ADA . However, 6 months after switching the clinical activity was lower in patients with previous ADA .When a subanalysis is performed taking only the group of patients with AS .No differences were observed in clinical activity and clinical improvement between patients treated with a second anti-TNF drug with a mAb or fusion protein (ASDAS after 6 months of switching: 2.47 ± 1.12 with mAb vs. 2.53 ± 1.11 with fusion protein, In this article we studied the role of immunity against a first anti-TNF drug in the short-term response to a second anti-TNF drug in a group of SpA patients. We show that patients in whom drug discontinuation was associated with ADA development achieved a better clinical response at 6 months after switching than patients who had not developed ADA.Anti-TNF drugs are the only biological therapies available to treat SpA patients with an inadequate response to conventional treatment, and their efficacy has been demonstrated in several randomised placebo-controlled studies -9. HowevOnly a few prospective studies have reported detailed information about switching to a second anti-TNF drug in AS patients ,40,41. OThe immunogenicity of biological therapies has been shown to influence secondary inefficacy in rheumatic diseases ,28,42-52Currently, there are only two reports that relate immunogenicity status to the first anti-TNF drug and its clinical response after switching to a second anti-TNF drug in RA patients ,22. BartOur study has several limitations. The number of patients is relatively small because all patients came from the same centre, and SpA patients are less likely than RA patients to discontinue anti-TNF drug ,41. HoweSimilar to RA, the failure to respond to a first anti-TNF drug due to the development of ADA predicts a better clinical response to a second biological treatment in SpA. The presence of ADA against the first anti-TNF drug is a determining factor for the response to a second anti-TNF drug. The study of the immunogenicity in biological treatment failure may help predict the response to a second biological treatment in SpA.Ada: adalimumab; AS: ankylosing spondylitis; ASDAS: Ankylosing Spondylitis Disease Activity Score; ADA: anti-drug antibodies; ELISA: enzyme-linked immunosorbent assay; Etn: etanercept; Gol: golimumab; IBD: inflammatory bowel disease; Ifx: infliximab; mAb: monoclonal antibody RA: rheumatoid arthritis; SpA: spondyloarthritis; TNF: tumour necrosis factor.AB has received fees from Roche, Schering-Plough, Wyeth, Abbott, BMS and UCB. EM-M is a consultant and a member of speakers' bureaus for Pfizer, MSD, UCB and Abbott. ChP, DP-S and LN have received speaker honoraria from Pfizer. All other authors declare that they have no competing interests.ChP, DP-S, EM-M and AB wrote the article. DP-S, SGC, MTL-C and ChP carried out the data collection and databases. AV, DP, LN, LL, SGC, AB, EM-M and ChP performed the clinical evaluation of patients. JD, SGC and ChP performed the statistical analysis. DP-S and FA performed the laboratory assays. All authors read and approved the final manuscript.